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Imaging the cell surface: argon sputtering to expose inner cell structures.

Gelsomina De Stasio¹, Bradley H Frazer, Marco Girasole

¹University of Wisconsin-Madison, Department of Physics and Synchrotron Radiation Center, Stoughton, Wisconsin 53589, USA. pupa@src.wisc.edu

Microscopy Research and Technique

|January 15, 2004

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We developed an argon sputtering technique to reveal internal cell structures for surface imaging microscopies. This method allows detailed visualization of organelles and cellular components without causing damage, enhancing microscopy capabilities.

Area of Science:

Cell Biology
Microscopy Techniques
Surface Science

Background:

Conventional microscopies like SEM, AFM, and X-PEEM are limited to surface imaging.
Internal cellular structures remain largely inaccessible to these surface-sensitive techniques.

Purpose of the Study:

To introduce a novel argon sputtering method for progressively exposing inner cell structures.
To enable detailed imaging of intracellular components using surface-sensitive microscopies without causing apparent damage.

Main Methods:

Argon sputtering was applied to progressively reveal internal cellular structures.
Sputtering time was varied to control the depth of exposure.
Confocal fluorescence microscopy, TEM, SEM, and X-PEEM were used for comparative imaging.

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Main Results:

The argon sputtering method successfully exposed intracellular structures including cytoskeleton, vesicles, mitochondria, nuclear membrane, and nucleoli.
Comparative imaging confirmed the accurate representation of reference intracellular structures like cytoskeleton fibers, adhesion complexes, and secretory vesicles.
The method demonstrated minimal apparent damage to the cellular structures.

Conclusions:

Argon sputtering is a valuable new tool for surface-sensitive microscopy.
This technique expands the applicability of techniques like SEM, AFM, and X-PEEM to study internal cell architecture.
It provides a method for correlative imaging of surface and subsurface cellular features.